Chimeric Vector Construction For Higher-plant Transformation

Balázs, Ervin and Bouzoubaa, Salait and Guilley, Hubert and Jonard, Gerard and Paszkowski, Jerzy and Richards, Ken (1985) Chimeric Vector Construction For Higher-plant Transformation. GENE, 40 (2-3). pp. 343-348. ISSN 0378-1119

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Abstract

A chimeric vector pKR612B 1 was developed containing the neomycin phosphotransferase (APH) gene from the Tn5 transposon under the control of the gene VI promoter of cauliflower mosaic vírus (CaMV), and was used to transform higher plánt protoplasts. Plasmid pDOBó 12, the parental vector of pKR612Bl, has two unique restriction sites, Smal and BamHI, positioned just downstream of the CaMV gene VI promoter sequence. These unique cloning sites can be used fór any kind of gene insertion intő this vector. Using the polyethylene glycol transformation procedure, a large number of turnip and tobacco protoplasts were transformed and proved to be resistant to kanamycin (Km). From tobacco protoplasts whole Km-resistant plants were regenerated and shown to contain the integrated foreign gene. APH activity was detected in both transformed calli and in regenerated plants. DNA from transformed clones was analysed by Southern biot hybridization, showing the presence of the Tn5-derived gene.

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